人血浆中麦考酚酸及其葡糖苷酸代谢物HPLC法测定

HPLC Determination of Mycophenolic Acid and Its Glucuronide Metabolite in Human Plasma

目的:建立HPLC测定人血浆中麦考酚酸(MPA)及其代谢物葡糖醛酸结合物(MPAG)浓度的方法。方法:以安定为内标,血浆经乙腈蛋白沉淀后直接进样。采用Zorbax Eclipse XDB C_(18)(150 mm×4.6 mm,5μm),柱温:30℃,流动相:甲醇-0.1%三氟乙酸(55:45,pH=2.2),流速:1.0 mL·min^(-1),检测波长215 nm。结果:血浆内源性杂质和常用合并用药对样品测定无干扰,MPA和MPAG血药浓度分别在0.5-40μG·mL^(-1)和2.5—200μg·mL^(-1)范围内线性关系良好。MPA和MPAG的日内、日间RSD均小于8%,样品提取后在24 h内及3次冻融后稳定性良好。结论:该法快速,简便,准确,可用于MPA和MPAG的体内分析及常规的治疗药物监测。

Objective: To establish a method for determining the concentrations of mycophenolic acid (MPA) and its glucuronide (MPAG) in human plasma. Method: Using diazepam as internal standard, MPA and MPAG in plasma samples were determined by HPLC with protein precipitation and the detector wavelength was set at 215 nm. HPLC analytical column was Zorbax Eclipse XDB C_(18) (150 mm×4.6 mm,5μm)at 30℃. The mobile phase consisted of a mixture methanol-0.1% trifluoroacetic acid (55: 45, pH=2.2) pumped at a flow rate of 1.0 mL·min^(-1). Results: The drug-free plasma did not interfere with the determination of drugs and internal standard. Calibration curves of MPA and MPAG were linear between 0.5-40μg·mL^(-1) and 2.5-200μg·mL^(-1), respectively. The precision of within-day and between-day was good. Conclusions: The method established in the paper can be used to determine the concentrations of MPA and MPAG in human plasma. Method can also be applied to the clinical pharmacy research in vivo.